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| Rapid authentication of Whitmania Pigra Whitman based on loop-mediated isothermal amplification technology |
| Hits 1364 Download times 658 Received:June 03, 2022 |
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| DOI
10.11656/j.issn.1673-9043.2022.05.20 |
| Key Words
Whitmania Pigra Whitman;loop-mediated isothermal amplification technology;molecular identification;rapid authentication |
| Author Name | Affiliation | E-mail | | ZHANG Huanyu | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
The Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | WANG Wenxiu | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
The Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | GAO Han | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
The Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | TAN Wei | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
The Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | YU Xiaolei | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
The Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | LI Zhenguo | YouBo Pharmaceutical Co. Ltd., Mudanjiang 157000, China | lzgjzt@vip.163.com | | TIAN Xiaoxuan | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
The Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | tian_xiaoxuan@tjutcm.edu.cn |
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| Abstract
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| [Objective] We aim to establish a loop-mediated isothermal amplification(LAMP) technique for the rapid differentiate of Whitmania Pigra Whitman from other closely related species.[Methods] We designed LAMP primers based on the mitochondrial cytochrome c oxidase I(COI) gene sequences of Whitmania Pigra Whitman. Subsequently, LAMP amplification of leech was performed at a constant temperature of 64℃ using the newly designed LAMP primer sets.[Results] After the reaction, ladder-like bands were observed in positive samples by electrophoresis, whereas control groups are produced without bands. After adding 0.2 μL 10 000×SYBR GreenⅠdye, the positive samples changed color from orange to green under both ultraviolet and natural light, while the control groups remained orange.[Conclusion] LAMP technology can be used for the rapid authentication of Whitmania Pigra Whitman, and provides technical support for the on-site identification of leeches in herbal market. |
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